Neuromuscular Alterations After Ankle Sprains: An Animal Model to Establish Causal Links After Injury

Context: The mechanisms that contribute to the development of chronic ankle instability are not understood. Investigators have developed a hypothetical model in which neuromuscular alterations that stem from damaged ankle ligaments are thought to affect periarticular and proximal muscle activity. However, the retrospective nature of these studies does not allow a causal link to be established. Objective: To assess temporal alterations in the activity of 2 periarticular muscles of the rat ankle and 2 proximal muscles of the rat hind limb after an ankle sprain. Design: Controlled laboratory study. Setting: Laboratory. Patients or Other Participants: Five healthy adult male Long Evans rats (age = 16 weeks, mass = 400.0 ± 13.5 g). Intervention(s): Indwelling fine-wire electromyography (EMG) electrodes were implanted surgically into the biceps femoris, medial gastrocnemius, vastus lateralis, and tibialis anterior muscles of the rats. We recorded baseline EMG measurements while the rats walked on a motor-driven treadmill and then induced a closed lateral ankle sprain by overextending the lateral ankle ligaments. After ankle sprain, the rats were placed on the treadmill every 24 hours for 7 days, and we recorded postsprain EMG data. Main Outcome Measure(s): Onset time of muscle activity, phase duration, sample entropy, and minimal detectable change (MDC) were assessed and compared with baseline using 2-tailed dependent t tests. Results: Compared with baseline, delayed onset time of muscle activity was exhibited in the biceps femoris (baseline = −16.7 ± 54.0 milliseconds [ms]) on day 0 (5.2 ± 64.1 ms; t4 = −4.655, P = .043) and tibialis anterior (baseline = 307.0 ± 64.2 ms) muscles on day 3 (362.5 ± 55.9 ms; t4 = −5.427, P = .03) and day 6 (357.3 ± 39.6 ms; t4 = −3.802, P = .02). Longer phase durations were observed for the vastus lateralis (baseline = 321.9 ± 92.6 ms) on day 3 (401.3 ± 101.2 ms; t3 = −4.001, P = .03), day 4 (404.1 ± 93.0 ms; t3 = −3.320, P = .048), and day 5 (364.6 ± 105.2 ms; t3 = −3.963, P = .03) and for the tibialis anterior (baseline = 103.9 ± 16.4 ms) on day 4 (154.9 ± 7.8 ms; t3 = −4.331, P = .050) and day 6 (141.9 ± 16.2 ms; t3 = −3.441, P = .03). After sprain, greater sample entropy was found for the vastus lateralis (baseline = 0.7 ± 0.3) on day 6 (0.9 ± 0.4; t4 = −3.481, P = .03) and day 7 (0.9 ± 0.3; t4 = −2.637, P = .050) and for the tibialis anterior (baseline = 0.6 ± 0.4) on day 4 (0.9 ± 0.5; t4 = −3.224, P = .03). The MDC analysis revealed increased sample entropy values for the vastus lateralis and tibialis anterior. Conclusions: Manually inducing an ankle sprain in a rat by overextending the lateral ankle ligaments altered the complexity of muscle-activation patterns, and the alterations exceeded the MDC of the baseline data

Long-Lasting Impairments in Quadriceps Mitochondrial Health, Muscle Size, and Phenotypic Composition Are Present After Non-Invasive Anterior Cruciate Ligament Injury

Introduction: Despite rigorous rehabilitation aimed at restoring muscle health, anterior cruciate ligament (ACL) injury is often hallmarked by significant long-term quadriceps muscle weakness. Derangements in mitochondrial function are a common feature of various atrophying conditions, yet it is unclear to what extent mitochondria are involved in the detrimental sequela of quadriceps dysfunction after ACL injury. Using a preclinical, non-invasive ACL injury rodent model, our objective was to explore the direct effect of an isolated ACL injury on mitochondrial function, muscle atrophy, and muscle phenotypic transitions. Methods: A total of 40 male and female, Long Evans rats (16-week-old) were exposed to non-invasive ACL injury, while 8 additional rats served as controls. Rats were euthanized at 3, 7, 14, 28, and 56 days after ACL injury, and vastus lateralis muscles were extracted to measure the mitochondrial respiratory control ratio (RCR; state 3 respiration/state 4 respiration), mitochondrial reactive oxygen species (ROS) production, fiber cross sectional area (CSA), and fiber phenotyping. Alterations in mitochondrial function and ROS production were detected using two-way (sex:group) analyses of variance. To determine if mitochondrial characteristics were related to fiber atrophy, individual linear mixed effect models were run by sex. Results: Mitochondria-derived ROS increased from days 7 to 56 after ACL injury (30–100%, P \u3c 0.05), concomitant with a twofold reduction in RCR (P \u3c 0.05). Post-injury, male rats displayed decreases in fiber CSA (days 7, 14, 56; P \u3c 0.05), loss of IIa fibers (day 7; P \u3c 0.05), and an increase in IIb fibers (day 7; P \u3c 0.05), while females displayed no changes in CSA or phenotyping (P \u3e 0.05). Males displayed a positive relationship between state 3 respiration and CSA at days 14 and 56 (P \u3c 0.05), while females only displayed a similar trend at day 14 (P = 0.05). Conclusion: Long-lasting impairments in quadriceps mitochondrial health are present after ACL injury and play a key role in the dysregulation of quadriceps muscle size and composition. Our preclinical data indicate that using mitoprotective therapies may be a potential therapeutic strategy to mitigate alterations in muscle size and characteristic after ACL injury

Corticospinal Tract Structure and Excitability in Patients with Anterior Cruciate Ligament Reconstruction: A DTI and TMS Study

© 2019 Background: Underlying neural factors contribute to poor outcomes following anterior cruciate ligament reconstruction (ACLR). Neurophysiological adaptations have been identified in corticospinal tract excitability, however limited evidence exists on neurostructural changes that may influence motor recovery in ACLR patients. Objective: To 1) quantify hemispheric differences in structural properties of the corticospinal tract in patients with a history of ACLR, and 2) assess the relationship between excitability and corticospinal tract structure. Methods: Ten participants with ACLR (age: 22.6 ± 1.9 yrs; height: 166.3 ± 7.5 cm; mass: 65.4 ± 12.6 kg, months from surgery: 70.0 ± 23.6) volunteered for this cross-sectional study. Corticospinal tract structure (volume; fractional anisotropy [FA]; axial diffusivity [AD]; radial diffusivity [RD]; mean diffusivity [MD]) was assessed using diffusion tensor imaging, and excitability was assessed using transcranial magnetic stimulation (motor evoked potentials normalized to maximal muscle response [MEP]) for each hemisphere. Hemispheric differences were evaluated using paired samples t-tests. Correlational analyses were conducted on structural and excitability outcomes. Results: The hemisphere of the ACLR injured limb (i.e. hemisphere contralateral to the ACLR injured limb) demonstrated lower volume, lower FA, higher MD, and smaller MEPs compared to the hemisphere of the non-injured limb, indicating disrupted white matter structure and a reduction in excitability of the corticospinal tract. Greater corticospinal tract excitability was associated with larger corticospinal tract volume. Conclusions: ACLR patients demonstrated asymmetry in structural properties of the corticospinal tract that may influence the recovery of motor function following surgical reconstruction. More research is warranted to establish the influence of neurostructural measures on patient outcomes and response to treatment in ACLR populations

Presentation1_Quadriceps muscle atrophy after non-invasive anterior cruciate ligament injury: evidence linking to autophagy and mitophagy.zip

Introduction: Anterior cruciate ligament (ACL) injury is frequently accompanied by quadriceps muscle atrophy, a process closely linked to mitochondrial health and mitochondria-specific autophagy. However, the temporal progression of key quadricep atrophy-mediating events following ACL injury remains poorly understood. To advance our understanding, we conducted a longitudinal study to elucidate key parameters in quadriceps autophagy and mitophagy.Methods: Long-Evans rats were euthanized at 7, 14, 28, and 56 days after non-invasive ACL injury that was induced via tibial compression overload; controls were not injured. Vastus lateralis muscle was extracted, and subsequent immunoblotting analysis was conducted using primary antibodies targeting key proteins involved in autophagy and mitophagy cellular processes.Results: Our findings demonstrated dynamic changes in autophagy and mitophagy markers in the quadriceps muscle during the recovery period after ACL injury. The early response to the injury was characterized by the induction of autophagy at 14 days (Beclin1), indicating an initial cellular response to the injury. Subsequently, at 14 days we observed increase in the elongation of autophagosomes (Atg4B), suggesting a potential remodeling process. The autophagosome flux was also augmented between 14- and 28 days (LC3-II/LC3-I ratio and p62). Notably, at 56 days, markers associated with the elimination of damaged mitochondria were elevated (PINK1, Parkin, and VDAC1), indicating a possible ongoing cellular repair and restoration process.Conclusion: These data highlight the complexity of muscle recovery after ACL injury and underscore the overlooked but crucial role of autophagy and mitophagy in promoting the recovery process.</p

Anterior Cruciate Ligament Research Retreat VIII Summary Statement: An Update on injury Risk Identification and Prevention Across The Anterior Cruciate Ligament injury Continuum, March 14-16, 2019, Greensboro, Nc

© by the National Athletic Trainers\u27 Association, Inc. Advances in research continue to shape what we know about the ACL injury continuum and the important directions for future research that are needed to move the field forward. Our expectation is that these proceedings will continue to stimulate strategic research initiatives to more effectively identify those at risk and promote high-quality clinical interventions to prevent the initial injury from occurring and to improve both the short-term and long-term patient outcomes when ACL injury does occur